Functional and phenotypic properties of peripheral T cells anergized by autologous CD3+ depleted bone marrow cells

Yide Jin, Laphalle Fuller, Manuel Carreno, Violet Esquenazi, Bonnie B Blomberg, Yu Tao Wei, Gaetano Ciancio, George W Burke, Andreas Tzakis, Camillo Ricordi, Joshua Miller

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

We have previously demonstrated that bone marrow cells (BMC) inhibit the generation of autologous Epstein-Barr virus (EBV) -specific cytotoxic T lymphocytes (CTL). It was also observed that CD3+ cells obtained after 7 days of culture in the presence of autologous BMC could be used as inhibitors of EBV-CTL generation. In the present study, we examined these BMC induced regulator CD3+ T cells with respect to phenotype, function, and T-cell activation pathways. We also questioned if the CD3+ regulatory cell function is mediated by their direct effect on peripheral T cells or on the ability of antigen presenting cells (APC) to stimulate peripheral T cells. To answer this, CD3+ cells from peripheral blood lymphocytes (PBL) were cultured with either CD3-depleted BMC or with CD3-depleted PBL. The CD3+ cells were then isolated with immunomagnetic beads, designated as TBM and TPBL, and were compared in functional studies. There was an increase in the expression of CD25 on TBM cells. The TBM cells also expressed less CD122 and a decreased number of CD3 molecules per cell. Both TBM and TPBL cell populations responded to mitogen (PHA) to the same magnitude. However, when stimulated through the CD3 complex with anti-CD3 monoclonal antibody (mAb), the TBM cells had a significantly decreased response than did TPBL. The addition of IL-2 to these latter cultures augmented, but could not fully restore, the response. Additionally, stimulation of TBM cells with allogeneic cells failed to produce cytotoxic T cells. These "anergized" TBM and "nonanergized" (control) TPBL cells were added as third-party cells to a CTL generating culture of autologous PBL stimulated with allogeneic cells. The TBM cells exhibited suppressor function and inhibited the generation of CTL, in contrast with TPBL. The effect of TBM cells on direct and indirect antigen presentation pathways demonstrated that TBM primarily effected indirect, but not direct, alloantigen presentation. To further explore the cytoplasmic T-cell activation events that occurred after the coculture of the PBL T cells with BMC, the levels of zeta-associated protein 70 (ZAP70) and extracellular receptor-activated kinase (ERK) were determined. There was a decrease in ZAP70 levels in the TBM, which correlated with its reduced expression of cell surface CD3 and the attenuated response to anti-CD3 mAb activation. However, the activity of ERK was equally expressed by TBM and TPBL. It, therefore, appears that the culturing of peripheral T cells with (non-T) BMC anergizes these cells (which become refractory to stimulation through the T-cell receptors), and induces immune suppressor function. These in vitro observations may provide a mechanism by which infused donor BMC serve to downregulate T-cell immunity.

Original languageEnglish
Pages (from-to)567-575
Number of pages9
JournalHuman Immunology
Volume63
Issue number7
DOIs
StatePublished - Jul 6 2002

Fingerprint

Bone Marrow Cells
T-Lymphocytes
Cytotoxic T-Lymphocytes
Lymphocytes
Human Herpesvirus 4
Phosphotransferases
Monoclonal Antibodies
CD3 Antigens
Isoantigens
Antigen Presentation
Antigen-Presenting Cells
Coculture Techniques
T-Cell Antigen Receptor
Mitogens
Interleukin-2
Immunity
Blood Cells
Proteins

Keywords

  • Anergy
  • Antigen presenting cells
  • Bone marrow
  • Infectious tolerance
  • Suppressor T cells

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Functional and phenotypic properties of peripheral T cells anergized by autologous CD3+ depleted bone marrow cells. / Jin, Yide; Fuller, Laphalle; Carreno, Manuel; Esquenazi, Violet; Blomberg, Bonnie B; Wei, Yu Tao; Ciancio, Gaetano; Burke, George W; Tzakis, Andreas; Ricordi, Camillo; Miller, Joshua.

In: Human Immunology, Vol. 63, No. 7, 06.07.2002, p. 567-575.

Research output: Contribution to journalArticle

Jin, Yide ; Fuller, Laphalle ; Carreno, Manuel ; Esquenazi, Violet ; Blomberg, Bonnie B ; Wei, Yu Tao ; Ciancio, Gaetano ; Burke, George W ; Tzakis, Andreas ; Ricordi, Camillo ; Miller, Joshua. / Functional and phenotypic properties of peripheral T cells anergized by autologous CD3+ depleted bone marrow cells. In: Human Immunology. 2002 ; Vol. 63, No. 7. pp. 567-575.
@article{6d67ee3c0c40414a8d2574af21cf4d4d,
title = "Functional and phenotypic properties of peripheral T cells anergized by autologous CD3+ depleted bone marrow cells",
abstract = "We have previously demonstrated that bone marrow cells (BMC) inhibit the generation of autologous Epstein-Barr virus (EBV) -specific cytotoxic T lymphocytes (CTL). It was also observed that CD3+ cells obtained after 7 days of culture in the presence of autologous BMC could be used as inhibitors of EBV-CTL generation. In the present study, we examined these BMC induced regulator CD3+ T cells with respect to phenotype, function, and T-cell activation pathways. We also questioned if the CD3+ regulatory cell function is mediated by their direct effect on peripheral T cells or on the ability of antigen presenting cells (APC) to stimulate peripheral T cells. To answer this, CD3+ cells from peripheral blood lymphocytes (PBL) were cultured with either CD3-depleted BMC or with CD3-depleted PBL. The CD3+ cells were then isolated with immunomagnetic beads, designated as TBM and TPBL, and were compared in functional studies. There was an increase in the expression of CD25 on TBM cells. The TBM cells also expressed less CD122 and a decreased number of CD3 molecules per cell. Both TBM and TPBL cell populations responded to mitogen (PHA) to the same magnitude. However, when stimulated through the CD3 complex with anti-CD3 monoclonal antibody (mAb), the TBM cells had a significantly decreased response than did TPBL. The addition of IL-2 to these latter cultures augmented, but could not fully restore, the response. Additionally, stimulation of TBM cells with allogeneic cells failed to produce cytotoxic T cells. These {"}anergized{"} TBM and {"}nonanergized{"} (control) TPBL cells were added as third-party cells to a CTL generating culture of autologous PBL stimulated with allogeneic cells. The TBM cells exhibited suppressor function and inhibited the generation of CTL, in contrast with TPBL. The effect of TBM cells on direct and indirect antigen presentation pathways demonstrated that TBM primarily effected indirect, but not direct, alloantigen presentation. To further explore the cytoplasmic T-cell activation events that occurred after the coculture of the PBL T cells with BMC, the levels of zeta-associated protein 70 (ZAP70) and extracellular receptor-activated kinase (ERK) were determined. There was a decrease in ZAP70 levels in the TBM, which correlated with its reduced expression of cell surface CD3 and the attenuated response to anti-CD3 mAb activation. However, the activity of ERK was equally expressed by TBM and TPBL. It, therefore, appears that the culturing of peripheral T cells with (non-T) BMC anergizes these cells (which become refractory to stimulation through the T-cell receptors), and induces immune suppressor function. These in vitro observations may provide a mechanism by which infused donor BMC serve to downregulate T-cell immunity.",
keywords = "Anergy, Antigen presenting cells, Bone marrow, Infectious tolerance, Suppressor T cells",
author = "Yide Jin and Laphalle Fuller and Manuel Carreno and Violet Esquenazi and Blomberg, {Bonnie B} and Wei, {Yu Tao} and Gaetano Ciancio and Burke, {George W} and Andreas Tzakis and Camillo Ricordi and Joshua Miller",
year = "2002",
month = "7",
day = "6",
doi = "10.1016/S0198-8859(02)00402-0",
language = "English",
volume = "63",
pages = "567--575",
journal = "Human Immunology",
issn = "0198-8859",
publisher = "Elsevier Inc.",
number = "7",

}

TY - JOUR

T1 - Functional and phenotypic properties of peripheral T cells anergized by autologous CD3+ depleted bone marrow cells

AU - Jin, Yide

AU - Fuller, Laphalle

AU - Carreno, Manuel

AU - Esquenazi, Violet

AU - Blomberg, Bonnie B

AU - Wei, Yu Tao

AU - Ciancio, Gaetano

AU - Burke, George W

AU - Tzakis, Andreas

AU - Ricordi, Camillo

AU - Miller, Joshua

PY - 2002/7/6

Y1 - 2002/7/6

N2 - We have previously demonstrated that bone marrow cells (BMC) inhibit the generation of autologous Epstein-Barr virus (EBV) -specific cytotoxic T lymphocytes (CTL). It was also observed that CD3+ cells obtained after 7 days of culture in the presence of autologous BMC could be used as inhibitors of EBV-CTL generation. In the present study, we examined these BMC induced regulator CD3+ T cells with respect to phenotype, function, and T-cell activation pathways. We also questioned if the CD3+ regulatory cell function is mediated by their direct effect on peripheral T cells or on the ability of antigen presenting cells (APC) to stimulate peripheral T cells. To answer this, CD3+ cells from peripheral blood lymphocytes (PBL) were cultured with either CD3-depleted BMC or with CD3-depleted PBL. The CD3+ cells were then isolated with immunomagnetic beads, designated as TBM and TPBL, and were compared in functional studies. There was an increase in the expression of CD25 on TBM cells. The TBM cells also expressed less CD122 and a decreased number of CD3 molecules per cell. Both TBM and TPBL cell populations responded to mitogen (PHA) to the same magnitude. However, when stimulated through the CD3 complex with anti-CD3 monoclonal antibody (mAb), the TBM cells had a significantly decreased response than did TPBL. The addition of IL-2 to these latter cultures augmented, but could not fully restore, the response. Additionally, stimulation of TBM cells with allogeneic cells failed to produce cytotoxic T cells. These "anergized" TBM and "nonanergized" (control) TPBL cells were added as third-party cells to a CTL generating culture of autologous PBL stimulated with allogeneic cells. The TBM cells exhibited suppressor function and inhibited the generation of CTL, in contrast with TPBL. The effect of TBM cells on direct and indirect antigen presentation pathways demonstrated that TBM primarily effected indirect, but not direct, alloantigen presentation. To further explore the cytoplasmic T-cell activation events that occurred after the coculture of the PBL T cells with BMC, the levels of zeta-associated protein 70 (ZAP70) and extracellular receptor-activated kinase (ERK) were determined. There was a decrease in ZAP70 levels in the TBM, which correlated with its reduced expression of cell surface CD3 and the attenuated response to anti-CD3 mAb activation. However, the activity of ERK was equally expressed by TBM and TPBL. It, therefore, appears that the culturing of peripheral T cells with (non-T) BMC anergizes these cells (which become refractory to stimulation through the T-cell receptors), and induces immune suppressor function. These in vitro observations may provide a mechanism by which infused donor BMC serve to downregulate T-cell immunity.

AB - We have previously demonstrated that bone marrow cells (BMC) inhibit the generation of autologous Epstein-Barr virus (EBV) -specific cytotoxic T lymphocytes (CTL). It was also observed that CD3+ cells obtained after 7 days of culture in the presence of autologous BMC could be used as inhibitors of EBV-CTL generation. In the present study, we examined these BMC induced regulator CD3+ T cells with respect to phenotype, function, and T-cell activation pathways. We also questioned if the CD3+ regulatory cell function is mediated by their direct effect on peripheral T cells or on the ability of antigen presenting cells (APC) to stimulate peripheral T cells. To answer this, CD3+ cells from peripheral blood lymphocytes (PBL) were cultured with either CD3-depleted BMC or with CD3-depleted PBL. The CD3+ cells were then isolated with immunomagnetic beads, designated as TBM and TPBL, and were compared in functional studies. There was an increase in the expression of CD25 on TBM cells. The TBM cells also expressed less CD122 and a decreased number of CD3 molecules per cell. Both TBM and TPBL cell populations responded to mitogen (PHA) to the same magnitude. However, when stimulated through the CD3 complex with anti-CD3 monoclonal antibody (mAb), the TBM cells had a significantly decreased response than did TPBL. The addition of IL-2 to these latter cultures augmented, but could not fully restore, the response. Additionally, stimulation of TBM cells with allogeneic cells failed to produce cytotoxic T cells. These "anergized" TBM and "nonanergized" (control) TPBL cells were added as third-party cells to a CTL generating culture of autologous PBL stimulated with allogeneic cells. The TBM cells exhibited suppressor function and inhibited the generation of CTL, in contrast with TPBL. The effect of TBM cells on direct and indirect antigen presentation pathways demonstrated that TBM primarily effected indirect, but not direct, alloantigen presentation. To further explore the cytoplasmic T-cell activation events that occurred after the coculture of the PBL T cells with BMC, the levels of zeta-associated protein 70 (ZAP70) and extracellular receptor-activated kinase (ERK) were determined. There was a decrease in ZAP70 levels in the TBM, which correlated with its reduced expression of cell surface CD3 and the attenuated response to anti-CD3 mAb activation. However, the activity of ERK was equally expressed by TBM and TPBL. It, therefore, appears that the culturing of peripheral T cells with (non-T) BMC anergizes these cells (which become refractory to stimulation through the T-cell receptors), and induces immune suppressor function. These in vitro observations may provide a mechanism by which infused donor BMC serve to downregulate T-cell immunity.

KW - Anergy

KW - Antigen presenting cells

KW - Bone marrow

KW - Infectious tolerance

KW - Suppressor T cells

UR - http://www.scopus.com/inward/record.url?scp=0035985429&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035985429&partnerID=8YFLogxK

U2 - 10.1016/S0198-8859(02)00402-0

DO - 10.1016/S0198-8859(02)00402-0

M3 - Article

VL - 63

SP - 567

EP - 575

JO - Human Immunology

JF - Human Immunology

SN - 0198-8859

IS - 7

ER -